Related Experiment Video
Updated: Jun 20, 2026
A 96 Well Microtiter Plate-based Method for Monitoring Formation and Antifungal Susceptibility Testing of Candida albicans Biofilms
Published on: October 21, 2010
Plate assay for fungal enzymes using cellophane membranes
Shu-Ying Liu1, Pei-Char Feng, Chaur-Tsuen Lo
1Department of Molecular Biotechnology, Da-Yeh University, Changhua 51591, Taiwan.
Abstract:
Fungal mycelia mass and pigments are major obstacles to investigating the secretion of bioactive substances such as enzyme activities using a plate assay. In this study, we applied a cellophane membrane and demonstrated that it can block mycelia mass and conidia (especially pigmented spores that would likely interfere with any subsequent color development-based activity detection) while allowing secreting enzymes to pass through. Visual observation after lifting the cellophane membrane and the collected mycelia and conidia indicated that the bioactivities on specific plates were improved significantly, although some fungal growth hurdle was noted. This proved to be true whether the assays were color development based or not.
Insights
This study introduces a cellophane membrane technique to improve the detection of fungal bioactive substances in plate assays. The method effectively blocks fungal growth, enhancing enzyme activity measurements.
Area of Science:
- Mycology
- Biochemistry
- Enzyme Assays
Background:
- Fungal mycelial mass and pigments impede the accurate measurement of secreted bioactive substances using plate assays.
- Detecting enzyme activities and other secreted compounds is crucial for understanding fungal biology and applications.
Purpose of the Study:
- To develop and validate a method using a cellophane membrane to overcome limitations in fungal plate assays.
- To improve the detection sensitivity of secreted bioactive substances, including enzyme activities.
Main Methods:
- A cellophane membrane was employed in plate assays to separate fungal biomass from the assay medium.
- The membrane's ability to block mycelia and pigmented conidia while allowing enzyme secretion was assessed.
- Bioactivity detection was performed with and without the cellophane membrane to compare results.
Main Results:
- The cellophane membrane successfully blocked fungal mycelia and pigmented spores, preventing interference with activity detection.
- Significant improvements in the detection of bioactivities were observed on the assay plates after using the membrane.
- The method proved effective for both colorimetric and non-colorimetric enzyme activity assays.
Conclusions:
- Cellophane membrane application is a viable strategy to enhance the investigation of secreted fungal enzymes and bioactive compounds.
- This technique offers a significant improvement for plate assay sensitivity, particularly in the presence of pigmented fungi.
- The method facilitates more accurate assessment of fungal metabolic secretions, aiding further research and application development.
